Research Article

Efficient isolation and chondrogenic differentiation of adult mesenchymal stem cells with fibrin microbeads and micronized collagen sponges

† Author for correspondence

Biotechnology & Radiobiology Laboratories, Sharett Institute of Oncology, Hadassah Hebrew University Medical Center, POB 12000, Jerusalem, Israel.

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Email the corresponding author at RAFI@hadassah.org.il

Published Online:8 Mar 2010https://doi.org/10.2217/rme.09.90

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Abstract

Background: Mesenchymal stem cells (MSCs) have been demonstrated to potentially undergo chondrogenic differentiation. We propose a new matrix for stem cell-based chondrogenesis using dense fibrin microbeads (FMBs) combined with grounded dehydrothermally crosslinked collagen sponges (micronized collagen). Methods: In this study, MSCs were isolated from bone marrow of transgenic green fluorescent protein C57/Bl mice by FMBs in high yield. After 48 h in slowly rotating suspension culture, micronized collagen was added. Results: The cells on the FMBs migrated to the collagen pieces and formed aggregates that developed into cartilage-like structures. Following chondrogenic differentiation, alcian blue staining and collagen type II immunohistochemistry demonstrated the presence of chondrocytes in the 3D structures. PCR for the expression of aggrecan and collagen type II genes supported these findings. The in vitro structures that formed were used for ectopic subdermal implantation in wild-type C57/Bl mice. However, the chondrogenic markers faded relative to the pre-implant in vitro structures. Conclusion: We propose that FMBs with micronized collagen could serve as a simple technology for MSC isolation and chondrogenesis as a basis for implantation.

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Published online 8 March 2010

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Acknowledgements

The authors wish to thank Dan Lesnoy from Forticell Biosciences Inc. (NY, USA) for the development and the preparation of the micronized collagen. We also thank Dimitry Belenky for his help in the production of the fibrin microbeads used in this study and Ezra Rachamin for his help in the sample preparations for scanning-electron microscopy readings.

Financial & competing interests disclosure

This work was partially supported by HAPTO Biotech Israel Ltd, a subdivision of Forticell Biosciences Inc. (NY, USA). The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

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